环状RNA-102231增加结肠癌HT-29细胞对顺铂耐药性的作用及其机制  

Effect of cyclic RNA-102231 on cisplatin resistance in HT-29 colon cancer cells and its mechanism

作　　者：王培培 周静 李铮[2] 李海[3] 李霄[3] Wang Peipei;Zhou Jing;Li Zheng;Li Hai;Li Xiao(Department of Clinical Pharmacy,Zhumadian Central Hospital,Zhumadian 463000,China;Department of General Surgery,Jiangsu Province Hospital,Nanjing 210000,China;Department of Pathology,Jiangsu Province Hospital,Nanjing 210OOO,China)

机构地区：[1]河南省驻马店市中心医院临床药学室,463000 [2]江苏省人民医院普通外科,南京210000 [3]江苏省人民医院病理学部,南京210000

出　　处：《中华普通外科杂志》2020年第7期558-562,共5页Chinese Journal of General Surgery

摘　　要：目的探讨环状RNA-102231(circRNA-102231)增加结肠癌HT-29细胞对顺铂耐药性的作用及其机制.方法用Western blot和RT-qPCR检测顺铂耐药的HT-29/DDP细胞株circRNA-102231和ABCB1的表达水平;在HT-29细胞中分别过表达或敲降circRNA-102231后加入不同浓度的顺铂,用CCK-8法检测细胞活力,Western blot检测ABCB1蛋白的表达水平,RT-qPCR检测circRNA-102231和ABCB1的mRNA表达水平;用数据库预测并验证与circRNA-102231相结合的micmRNA;激活或抑制ERK的同时过表达miR-145,Western blot和RT-qPCR检测ABCB1的蛋白和mRNA表达水平,荧光素酶报告基因检测ABCB1启动子活性.结果与HT-29细胞相比,HT-29/DDP细胞中circRNA-102231和ABCB1的表达水平均显著上调(均P<0.05);过表达circRNA-102231能显著上调ABCB1的表达水平,降低细胞对顺铂的敏感性(P<0.05);敲降circRNA-102231能下调ABCB1的表达水平,增加细胞对顺铂的敏感性(P<0.05);circRNA-102231可以招募miR-145并能够促进其表达;miR-145能促进ERK磷酸化,激活或抑制ERK活性能够促进或上调ABCB1的表达水平和启动子活性(P<0.05).结论circRNA-102231可能通过miR-145/ERK通路上调ABCB1的表达并提高HT-29细胞对顺铂的耐药性.Objective To investigate the effect of circRNA-102231 on the sensitivity of colon cancer HT-29 cells to cisplatin and its mechanism.Methods The expression of circRNA-102231 and ABCBl were detected by Western blot and RT-qPCR in cisplatin-resistant HT-29/DDP cell lines.Overexpress or knockdown circRNA-102231 in HT-29 cells were respectively followed by treatment with cisplatin,CCK-8 method was used to detect cell viability,Western blot and RT-qPCR were used to detect the protein and mRNA expression levels of circ-102231 and ABCBl;The circRNA-102231 combined microRNA was predicted by the database and verified with RT-qPCR;Activation or inhibition of ERK and overexpression of miR-145,Western blot and RT-qPCR were used to detect the protein and mRNA levels of ABCBl,and the luciferase reporter assay was used to detect the ABCBl promoter activity.Results Compared with HT-29 cells,the expression levels of circRNA-102231 and ABCBl were significantly up-regulated in HT-29/DDP cells(all P<0.05).Overexpression of circRNA-102231 up-regulated the expression of ABCBl and decreased the sensitivity of cells to cisplatin(P<0.05).Knockdown of circRNA-102231 down-regulated the expression of ABCBl,hence increased the sensitivity to cisplatin(P<0.05);circRNA-102231 can recruit miR-145 and promote its expression;miR-145 induce ERK phosphorylation,activation or inhibition of ERK activity can promote or up regulate the expression level and promote activity of ABCBl(P<0.05).Conclusion circRNA-102231 may increase the resistance of HT-29 cells to cisplatin by promoting the up-regulation of ABCBl through miRNA-145/ERK pathway.

关 键 词：结肠肿瘤 多药耐药相关蛋白质类 环状RNA 

分 类 号：R965[医药卫生—药理学]